Difference between revisions of "Team:Paris Saclay/Notebook/August/29"
MahnaziGEM (Talk | contribs) (Created page with "PCR sur colonie ====PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid==== ''Mahnaz'' * 2.5 µL DreamTaq Buffer * 0.5 µL of dNTPs (...") |
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| − | + | {{Team:Paris_Saclay/notebook_header}} | |
| + | = Monday 29<sup>th</sup> August= | ||
| + | |||
| + | ===Visualization=== | ||
====PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid==== | ====PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid==== | ||
| − | ''Mahnaz'' | + | ''By Mahnaz'' |
* 2.5 µL DreamTaq Buffer | * 2.5 µL DreamTaq Buffer | ||
| Line 25: | Line 28: | ||
|30 sec | |30 sec | ||
|- | |- | ||
| − | | | + | |52 |
|30 sec | |30 sec | ||
|- | |- | ||
| − | | | + | |72°C |
| − | | | + | |1 min |
|- | |- | ||
|Final Extension | |Final Extension | ||
| Line 37: | Line 40: | ||
|Hold | |Hold | ||
|4°C | |4°C | ||
| − | | | + | |\infinity\ |
|} | |} | ||
| − | + | [[Team:Paris_Saclay/Experiments#primers|Primers]] used were: | |
{| class="wikitable" | {| class="wikitable" | ||
| Line 48: | Line 51: | ||
|- | |- | ||
|Primers | |Primers | ||
| − | | | + | |pJET R and F |
|- | |- | ||
|Tm | |Tm | ||
| − | | | + | |60°C |
|- | |- | ||
|t | |t | ||
| Line 66: | Line 69: | ||
!Expected band size (bp) | !Expected band size (bp) | ||
|- | |- | ||
| − | | | + | |FRB in pJET |
| − | | | + | |474 |
|} | |} | ||
| − | [[File:T-- | + | [[File:T--Paris_saclay--2908.extension.png|400px|thumb|center|Result of migration ]] |
| + | The expected size was 474 pb and the 4 plasmids were sent to sequencing. | ||
| + | |||
| + | ====Samples preparation for sequencing==== | ||
| + | ''by Mahnaz'' | ||
| + | |||
| + | 15 µL of plasmides plasmids coding FRB from colonies 4,5,9 and 12 were sent to be sequenced. 20 µL of the primers pJET F (5µM) and pJET R (5µM) were sent for sequencing. | ||
| + | |||
| + | ====NanoDrop Measurements==== | ||
| + | ''By Mahnaz'' | ||
| + | |||
| + | {| class="wikitable" | ||
| + | !Sample | ||
| + | !Concentration (ng/µL) | ||
| + | |- | ||
| + | |PCR fragment FRB clone 4<div id="PCR fragment FRB clone 4"></div> | ||
| + | |58.43 | ||
| + | |- | ||
| + | |PCR fragment FRB clone 5<div id="PCR fragment FRB clone 5"></div> | ||
| + | |320.75 | ||
| + | |- | ||
| + | |PCR fragment FRB clone 9<div id="PCR fragment FRB clone 9"></div> | ||
| + | |830.99 | ||
| + | |- | ||
| + | |PCR fragment FRB clone 12<div id="PCR fragment FRB clone 12"></div> | ||
| + | |473.37 | ||
| + | |- | ||
| + | |} | ||
| − | + | {{Team:Paris_Saclay/notebook_footer}} | |
Latest revision as of 17:00, 9 October 2016
